New microscopy technologies will be developed and made available to biomedical researchers to bridge understanding of biological systems across gross anatomical and molecular scales. Project technology development efforts take on two important challenges: addressing the mesoscale range - from a few nanometers to 100 microns, required to see complexes in the context of cellular aggregates in tissues - and developing/advancing capabilities to enable multimodal imaging, that is, correlated imaging across a range of instruments (and their varying capabilities), to allow combining of data from the same specimens, providing new understanding of processes related to disease mechanisms. To achieve these two goals, activities will be in four related technology thrusts: probe development in Core 1, specimen development in Core 2, imaging instrument development in Core 3, and development of software tools to refine, integrate, analyze, and share microscopy data in Core 4. Core 1 will focus on labeling strategies for various new genetic or enzymatic probes. Core 2 will refine specimen-preparation protocols and materials to provide more suitable samples for examination. The combined outcome of these first two cores will be to streamline imaging using correlated microscopy by creating specimens better suited to a range of imaging modalities and enhancing the palette of in-situ markers to facilitate tracking areas of interest in related images and achieve their co-registration. Tracking methods, to be developed by activities of Cores 2 and 3, will be used to connect data in databases using new data analysis tools of Core 4, developing new tools for segmentation and annotation of images. Core 3 advances the imaging capabilities of multiple microscope types to deliver information not only with greater sensitivity and accuracy but more rapidly and seamlessly across scales. The work of all Cores, but especially the instrument development Core 3, will enable more sophisticated use of a next generation microscopes and their application to important biomedical research challenges. Core 4 develops software tools and infrastructure to refine, integrate, quantify, interpret, and add value to image data derived from the new instruments. The coordinated work among the four cores and collaborator projects will enable biomedical researchers to image wider, see deeper, increase contrast and differentiate complex structures, and observe complex phenomena at higher resolution and at faster time scales. Thus we will propel biomedical research requiring traversal of now difficult to navigate spatial scales, thereby facilitating new understanding of the molecular mechanisms underlying disease processes.